The goal of this research is to optimize in vitro and in vivo properties of Ontogen's mechanistically unique, novel, small molecule inhibitors of fructose 1,6-bisphosphatase (FBPase) for use in regulating glucose levels in diabetics. Previously discovered leads will be optimized for drug-like properties, safety, efficacy, and oral bioavailability. The successful compound will ultimately enter clinical development as a novel treatment for type 2 diabetes. Elevated blood glucose is a major cause of complications in diabetics. FBPase, a key enzyme in the production of glucose via gluconeogenesis, represents an important therapeutic target. Potent, non-carbohydrate, non-nucleoside FBPase inhibitors (IC50's < 10 microM) were identified by high-throughput screening of Ontogen's combinatorial libraries. Cocrystallization studies revealed the inhibitors bind a novel site at the interface of the FBPase homotetramer, preventing reorganization of loops critical for enzymatic activity. High-resolution structural information and experimental/computed physicochemical parameters will guide optimization. Focused combinatorial libraries, synthesized using proprietary OntoBLOCK(TM) instrumentation and purified with Ontogen's state-of-the-art OntoCHROM(TM) system, will be screened against FBPase. Cell-based assays will confirm that compounds inhibit gluconeogenesis. In vitro predictors of permeability, bioavailability, and stability will prioritize compounds for in vivo testing. Finally, preclinical toxicology, metabolism and pharmacokinetics will identify a development candidate for clinical trials. PROPOSED COMMERCIAL APPLICATION: Because of the increasing number of people suffering from diabetes in the US and worldwide, and a relative paucity of treatment options (see Research Plan, "Significance"), the need for new treatments for diabetes is critical, particularly for type 2 diabetes. The research we propose will build on our earlier discoveries and is expected to identify a clinical development candidate that will reduce excess glucose production via inhibition of fructose-1,6- bisphosphatase. This will provide a novel approach for glucose maintenance in the treatment of diabetes.